Integrated GC–MS- and LC–MS-Based Untargeted Metabolomics Studies of the Effect of Vitamin D3 on Pearl Production Traits in Pearl Oyster Pinctada fucata martensii

Abstract

Pearl oyster Pinctada fucata martensii is widely recognized for biomineralization and has been cultured for high-quality marine pearl production. To ascertain how dietary vitamin D3 (VD3) levels affect the features of pearl production by P. f. martensii and discover the mechanisms regulating this occurrence, five experimental diets with variable levels of VD3 were used with inclusion levels of 0, 500, 1,000, 3,000, and 10,000 IU/kg. The distinct inclusion levels were distributed into five experimental groups (EG1, EG2, EG3, EG4, and EG5). All the experimental groups were reared indoors except the control group (CG) reared at the sea. Pearl oysters, one year and a half old, were used in the grafting operation to culture pearls. During the growing period that lasted 137 days, EG3 had the highest survival rate, retention rate, and high-quality pearl rate. A similar trend was found for EG3 and CG with significantly higher pearl thickness and nacre deposition rates than other groups, but no significant differences were observed between them. A metabolomics profiling using GC–MS and LC–MS of pearl oysters fed with low quantities of dietary VD3 and optimal levels of dietary VD3 revealed 135 statistically differential metabolites (SDMs) (VIP > 1 and p < 0.05). Pathway analysis indicated that SDMs were involved in 32 pathways, such as phenylalanine metabolism, histidine metabolism, glycerophospholipid metabolism, alanine aspartate and glutamate metabolism, arginine and proline metabolism, glycerolipid metabolism, amino sugar and nucleotide sugar metabolism, and tyrosine metabolism. These results provide a theoretical foundation for understanding the impacts of VD3 on pearl production traits in pearl oyster and reinforce forthcoming prospects and application of VD3 in pearl oyster in aquaculture rearing conditions.

Dietary Carbohydrate and Protein Levels Affect the Growth Performance of Juvenile Peanut Worm (Sipunculus nudus): An LC–MS-Based Metabolomics Study

“…Data were scaled and logarithmically transformed to minimize the effect of noise and high variance among the variables. Data were analyzed as previously described (Yang et al, 2021). In the OPLS-DA analysis, the variable importance in projection (VIP) of the first principal component in OPLS-DA analysis was obtained.…”

“…A 100-µL sample was pipetted into an Eppendorf tube, and 400 µL of extract solution (acetonitrile: methanol = 1: 1) containing an isotopically labeled internal standard mixture was added. Metabolite extraction followed the methods of Yang et al (2021).…”

“…A UHPLC system (Vanquish, Thermo Fisher Scientific) with a UPLC BEH Amide column (2.1 mm × 100 mm, 1.7 µm) and a Q Exactive HFX mass spectrometer (Orbitrap MS, Thermo) was used for LC-MS/MS analyses. The analysis was carried out with an elution gradient as described by Yang et al (2021). The flow rate was 0.5 mL/min; the auto-sampler temperature was 4 • C; the column temperature was 30 • C; and the injection volume was 2 µL.…”

“…Metabolomics provides a "snapshot" of the metabolites and is particularly suitable for metabolic studies (Cappello et al, 2017(Cappello et al, , 2018. It is commonly used to quantify low-molecular-weight metabolites, such as sugars, lipids, and amino acids, through various tools such as gas chromatography-mass spectrometry, nuclear magnetic resonance, and liquid chromatography-mass spectrometry (LC-MS; Venter et al, 2018;Yang et al, 2021;Zhang J. B. et al, 2021).…”

Combined transcriptome and metabolite profiling analyses provide insights into the chronic toxicity of carbaryl and acetamiprid to Apis mellifera larvae

“…The metabolites extraction of the developing larvae was carried out as previously described 76 . The metabolite identification and quantification were performed using an UHPLC system (1290, Agilent Technologies) coupled to TripleTOF 5600 (Q-TOF, AB Sciex) at Shanghai Biotree biotech Co., Ltd. ( http://biotree.cn/index_en ) following their standard procedures and previously described by Yang et al 77 . Metabolite profiles were determined in five control and five treatment groups with each sample consisting of 10 homogenised larvae.…”